CN109507163A - H in a kind of aqueous solution2PO4Detection method - Google Patents
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- CN109507163A CN109507163A CN201811565770.5A CN201811565770A CN109507163A CN 109507163 A CN109507163 A CN 109507163A CN 201811565770 A CN201811565770 A CN 201811565770A CN 109507163 A CN109507163 A CN 109507163A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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Abstract
The invention discloses H in a kind of aqueous solution2PO4 ‑Detection method, be the super molecular complex that is prepared using cucurbit(7)uril and Hoechst-33258 as fluorescence probe, with the fluorescence probe to the H in water2PO4 ‑It is detected.Method of the invention has testing cost low, easy to operate, time-consuming short, and detects sensitive feature.
Description
Technical field
The present invention relates to a kind of H2PO4 -Detection method, H in especially a kind of aqueous solution2PO4 -Detection method.
Background technique
Dihydrogen phosphate ions are the important components of life system, play signal transduction in some life process
With the effect of energy storage.Therefore, have great importance to the detection of dihydrogen phosphate ions.
Hoechst is the compound of German Hoechst AG company synthesis, is dissolved in water and organic solvent.It is mainly used for living thin
Born of the same parents' label.The fluorescence intensity of Hoechst dyestuff enhances as pH value of solution increases.The reagent of all of which is all ordered with digital number
Name, that is to say, that Hoechst-33258 is the 33258th kind of compound of the said firm's synthesis.
Reported detection H2PO4 -Most common method is the chromatography of ions and electrode potential titration, these methods are logical
Accurate equipment, and sample treatment trouble are often needed, testing cost is high, and it is cumbersome, it takes a long time.Therefore, it designs a kind of low
Cost, easily simple, H in time-consuming short water2PO4 -Detection method be of great significance.And fluorescent spectrometry be able to solve it is above-mentioned
Problem, still, the super molecular complex being prepared using cucurbit(7)uril with Hoechst-33258 are detected as fluorescence probe
H in aqueous solution2PO4 -Method have not been reported.
Summary of the invention
The object of the present invention is to provide H in a kind of aqueous solution2PO4 -Detection method.Method of the invention has detection
It is at low cost, it is easy to operate, it is time-consuming short, and detect sensitive feature.
A kind of technical solution of the present invention: H in aqueous solution2PO4 -Detection method, be with cucurbit(7)uril and Hoechst-
33258 super molecular complexes being prepared are as fluorescence probe, with the fluorescence probe to the H in water2PO4 -It is detected.
H in aqueous solution above-mentioned2PO4 -Detection method, the specific steps are as follows:
1) water to be measured is taken, water to be measured is adjusted in alkalinity, obtains A product；
2) A product are added in fluorescence probe solution, obtain B product；
3) after B product are stored at room temperature 10-20min, to B product carry out fluorescence emission spectrum measurement, according to measurement result judge to
It surveys in water and whether contains H2PO4 -。
H in aqueous solution above-mentioned2PO4 -Detection method, in the step 1), using the sodium hydroxide of pH=11 adjust to
Water is surveyed in alkalinity.
H in aqueous solution above-mentioned2PO4 -Detection method, in the step 2), the preparation method of the fluorescence probe solution
It is as follows: cucurbit(7)uril and Hoechst-33258 are blended and dissolved in sodium hydroxide solution.
H in aqueous solution above-mentioned2PO4 -Detection method, the pH=11 of the sodium hydroxide solution.
H in aqueous solution above-mentioned2PO4 -Detection method, cucurbit(7)uril and Hoechst- in the fluorescence probe solution
33258 molar concentration rate is 2:1.
H in aqueous solution above-mentioned2PO4 -Detection method, in the fluorescence probe solution, the molar concentration of fluorescence probe is
1.00×10-4mol/L。
H in aqueous solution above-mentioned2PO4 -Detection method, in the step 3), the fixed excitation of fluorescence emission spectrum measurement
Wavelength is 353nm, and the method for measurement result judgement is, if B product fluorescence emission spectrum corresponds to the increasing of the fluorescent emission intensity at 475nm
By force, then illustrate to contain H in prepare liquid2PO4 -Do not increase if B product fluorescence emission spectrum corresponds to the fluorescent emission intensity at 475nm
By force, then illustrate in prepare liquid without H2PO4 -。
Beneficial effects of the present invention
1, the present invention prepares fluorescence probe using cucurbit(7)uril and Hoechst-33258 as raw material, passes through fluorescent spectrometry
Whether to contain H in prepare liquid2PO4 -It is measured, reduces testing cost.
2, the present invention only needs for prepare liquid to be adjusted to alkalinity, then instills in probe solution, is by analysis of spectral method
Can, sample making is simple, convenient for operation.
3, the present invention can intuitively determine result by the strong and weak variation of fluorescence spectrum, not need prolonged
Reaction and detection, substantially reduce the time of detection.
4, the present invention is minimum can be to H2PO4 -Concentration is 3.92 × 10-6Solution detected, have detection sensitive excellent
Point.
Detailed description of the invention
Attached drawing 1 is the chemical structure schematic diagram of fluorescence probe of the present invention；
Attached drawing 2 is fluorescence spectra of fluorescence probe in the presence of different anions；
Attached drawing 3 is the fluorescence spectra and canonical plotting (excitation of fluorescence probe solution fluorescence spectroscopic methodology detection H2PO4-
Wavelength is 353nm, slit 5nm, voltage 520v)；
Attached drawing 4 is interference spectrogram of the interfering ion to system.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated, but is not intended as the foundation limited the present invention.
The embodiment of the present invention
A kind of embodiment 1: H in aqueous solution2PO4 -Detection method, steps are as follows:
1) preparation of fluorescence probe solution: cucurbit(7)uril and Hoechst-33258 are mixed according to molar ratio 2:1, and molten
Solution prepares the dense of the super molecular complex of cucurbit(7)uril and Hoechst-33258 composition in the sodium hydroxide solution of pH=11
Degree is 1.00 × 10-4The fluorescence probe solution of mol/L；
2) water sample processing to be measured: taking water to be measured, water to be measured be adjusted to alkalinity using the sodium hydroxide of pH=11, standby
With；
3) it detects: the water sample to be measured that step 2) is adjusted to alkalinity is added dropwise to fluorescence probe solution made from step 1)
In, it is stored at room temperature after 10min to fix excitation wavelength 353nm to fluorescence probe solution and carry out fluorescence spectrometry, and observe glimmering
Light probe solution fluorescence Strength Changes illustrate to be measured if fluorescence emission spectrum corresponds to the enhancing of the fluorescent emission intensity at 475nm
Contain H in liquid2PO4 -If fluorescence emission spectrum, which corresponds to the fluorescent emission intensity at 475nm, not to be enhanced, illustrate in prepare liquid
Without H2PO4 -。
A kind of embodiment 2: H in aqueous solution2PO4 -Detection method, steps are as follows:
1) preparation of fluorescence probe solution: cucurbit(7)uril and Hoechst-33258 are mixed according to molar ratio 2:1, and molten
Solution prepares the dense of the super molecular complex of cucurbit(7)uril and Hoechst-33258 composition in the sodium hydroxide solution of pH=11
Degree is 1.00 × 10-4The fluorescence probe solution of mol/L；
2) water sample processing to be measured: taking water to be measured, water to be measured be adjusted to alkalinity using the sodium hydroxide of pH=11, standby
With；
3) it detects: the water sample to be measured that step 2) is adjusted to alkalinity is added dropwise to fluorescence probe solution made from step 1)
In, it is stored at room temperature after 20min to fix excitation wavelength 353nm to fluorescence probe solution and carry out fluorescence spectrometry, and observe glimmering
Light probe solution fluorescence Strength Changes illustrate to be measured if fluorescence emission spectrum corresponds to the enhancing of the fluorescent emission intensity at 475nm
Contain H in liquid2PO4 -If fluorescence emission spectrum, which corresponds to the fluorescent emission intensity at 475nm, not to be enhanced, illustrate in prepare liquid
Without H2PO4 -。
Experimental example
Method is as follows:
(1) preparation of the NaOH aqueous solution of pH=11:
The 1MNaOH solution for accurately pipetting 1.00mL, with secondary distilled water constant volume, is configured in the volumetric flask of 1000mL
The NaOH aqueous solution of pH=11；
(2) preparation of fluorescence probe standard solution:
26.84mgQ [7] and 5.34mgH33258 is accurately weighed, with the NaOH aqueous dissolution of pH=11 and is settled to
100mL.It obtains pH=11, Q [7] and H33258 molar concentration rate is 2:1, molar concentration is 1.00 × 10-4The fluorescence of mol/L
Probe standard solution；
(3) preparation of tetrabutyl ammonium hydrogen phosphate standard solution:
678.90mg tetrabutyl ammonium hydrogen phosphate is accurately weighed, with the NaOH aqueous dissolution of pH=11 and is settled to 10mL,
Up to pH=11, the H that molar concentration is 0.20mol/L2PO4 -Standard solution；
(4) preparation of other solions:
Analytically pure various 4-butyl ammoniums are accurately weighed, with the NaOH aqueous dissolution of pH=11, pH=11 is prepared, rubs
Other each anion standard solution that your concentration is 0.20mol/L；Accurately weigh required various perchlorate metal ions
Pure standard items are analyzed, with the NaOH aqueous dissolution of pH=11, it is each for other of 0.20mol/L to prepare pH=11, molar concentration
Metal ion standard solution；
(5) drafting of standard curve:
7,10mL volumetric flask are taken, every bottle is added 1.0 × 10-4It is accurate respectively to add after 1000 μ L of mol/L fluorescence probe solution
Enter 0.20mol/LH2PO4 -0 μ L of standard solution, 10 μ L, 20 μ L, 30 μ L, 40 μ L, 50 μ L, 60 μ L, it is water-soluble with the NaOH of pH=11
Liquid constant volume, shakes up, and after being placed at room temperature for 10-20min, fixed excitation wavelength 353nm carries out fluorescence emission spectrum measurement, with H2PO4 -
Concentration is abscissa, fluorescence probe emissive porwer (I at 475nm0) and addition various concentration H2PO4 -Fluorescent emission intensity (I)
Difference (Δ I) be ordinate, obtain standard curve.By the standard deviation of 20 blank solutions of slope and measurement of standard curve
Difference calculates probe Q [7]/H33258 and detects H2PO4 -Detection limit.Correlation analysis parameter is listed in table 1.
(6) anti-interference measurement:
Other anion (F is added in the alkaline aqueous solution of probe-, Cl-, Br-, I-, PF6 -, BF4 -, ClO2 -, ClO4 -,
SO4 2-, HSO4 -, HSO3 -, CO3 2-, CH3COO-, NO3 -, H2PO4 -, OH-) after, comparison fluorescent emission intensity (Fig. 2) shows the fluorescence
System selectivity with higher.H is added within the probe2PO4 -Afterwards, solution fluorescence emission spectrum corresponds to fluorescent emission at 497nm
Enhanced strength (Fig. 3), to establish a kind of H2PO4 -Detection method；Interfering ion (Li is added in system+, Na+, K+, Mg2+,
Ca2+, Ba2+, Al3+, Pb2+, Cu2+, Fe3+, Fe2+, Mn2+, Cl-, Br-, I-, OH-, NO3 -, CO3 2-, SO4 2-, ClO4 -) after, it compares glimmering
The experiment (Fig. 4) of light emitting intensity shows that the fluorescence system has certain anti-interference ability；
(7) sample detection:
Laboratory tap water is taken to take 0.2 μm of water phase membrane filtration of upper solution after the centrifugation of filter paper filtering centrifuge, add
Entering a small amount of alkali adjusting makes the pH=11 of water sample as prepare liquid, detects water sample, does not find there is H in water sample2PO4 -Presence；It takes
The volumetric flask of 10mL is several, on the basis of this, is separately added into certain density H using standard addition method2PO4 -Standard solution,
Make its concentration range in 0-12 × 10-4The fluorescence probe solution 1mL of pH=11 is added in mol/L, then uses the NaOH water of pH=11
Solution constant volume (volume of water sample >=50%), shakes up, and after being placed at room temperature for 10-20min, fixed excitation wavelength 353nm carries out fluorescence hair
Spectroscopic assay is penetrated, is measured in parallel 3 times, is averaged, standard recovery rate is tested according to IUPAC law regulation.Testing result is listed in table
2。
1 Q of table [7]/H33258 detects H2PO4 -Analysis parameter
| Ion | Calibration curve | R2 | Detection limit (mol/L) |
| H2PO4 - | △ F=3.60C-7.35 | 0.993 | 3.92×10-6 |
2 sample detection result of table
| Additional amount (× 10-5mol/L) | Measured amount (× 10-5mol/L) | The rate of recovery (%) |
| 20.00 | 19.96 | 99.80 |
| 60.00 | 60.14 | 100.23 |
| 100.00 | 98.17 | 98.17 |
The above described is only a preferred embodiment of the present invention, being not intended to limit the present invention in any form, appoint
What is to the above embodiments according to the technical essence of the invention any simply to repair without departing from technical solution of the present invention content
Change, equivalent variations and modification, all of which are still within the scope of the technical scheme of the invention.
Claims (8)
1. H in a kind of aqueous solution2PO4 -Detection method, it is characterised in that: be to be prepared with cucurbit(7)uril and Hoechst-33258
Obtained super molecular complex is as fluorescence probe, with the fluorescence probe to the H in water2PO4 -It is detected.
2. H in aqueous solution according to claim 12PO4 -Detection method, which is characterized in that specific step is as follows:
1) water to be measured is taken, water to be measured is adjusted in alkalinity, obtains A product；
2) A product are added in fluorescence probe solution, obtain B product；
3) after B product are stored at room temperature 10-20min, fluorescence emission spectrum measurement is carried out to B product, water to be measured is judged according to measurement result
In whether contain H2PO4 -。
3. H in aqueous solution according to claim 22PO4 -Detection method, it is characterised in that: in the step 1), use
The sodium hydroxide of pH=11 adjusts water to be measured in alkalinity.
4. H in aqueous solution according to claim 1 or 22PO4 -Detection method, it is characterised in that: in the step 2),
The fluorescence probe solution the preparation method is as follows: cucurbit(7)uril and Hoechst-33258 are blended and dissolved in sodium hydroxide
In solution.
5. H in aqueous solution according to claim 42PO4 -Detection method, it is characterised in that: the sodium hydroxide solution
PH=11.
6. H in aqueous solution according to claim 22PO4 -Detection method, it is characterised in that: the fluorescence probe solution
The molar concentration rate of middle cucurbit(7)uril and Hoechst-33258 are 2:1.
7. H in aqueous solution according to claim 22PO4 -Detection method, it is characterised in that: the fluorescence probe solution
In, the molar concentration of fluorescence probe is 1.00 × 10-4mol/L。
8. H in aqueous solution according to claim 22PO4 -Detection method, it is characterised in that: in the step 3), fluorescence
The fixation excitation wavelength of emission spectrometry is 353nm, and the method for measurement result judgement is, if B product fluorescence emission spectrum is corresponding
Fluorescent emission intensity enhancing at 475nm, then illustrate to contain H in prepare liquid2PO4 -If B product fluorescence emission spectrum corresponds to 475nm
The fluorescent emission intensity at place does not enhance, then illustrates in prepare liquid without H2PO4 -。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113150770A (en) * | 2021-02-24 | 2021-07-23 | 武汉纺织大学 | Molecular assembly fluorescent probe and preparation method and application thereof |
| CN113670867A (en) * | 2021-07-16 | 2021-11-19 | 贵州大学 | Supermolecule fluorescence sensing array and method for detecting multiple metal ions in aqueous solution |
| CN113880851A (en) * | 2021-11-12 | 2022-01-04 | 广东科学技术职业学院 | Trifluorene bridged hexaimidazole macrocyclic compound and preparation method and application thereof |
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| CN106198469A (en) * | 2016-06-29 | 2016-12-07 | 南京大学 | A kind of utilize Hoechst 33258 fluorescent dye to the detection method of active o content in living cells |
| CN106905958A (en) * | 2017-03-21 | 2017-06-30 | 贵州大学 | A kind of fluorescence probe based on trans cucurbit(7)uril, preparation method and application |
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2018
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| EP1373889A2 (en) * | 2000-07-31 | 2004-01-02 | Maxygen, Inc. | Biosensors, reagents and diagnostic applications of directed evolution |
| CN106198469A (en) * | 2016-06-29 | 2016-12-07 | 南京大学 | A kind of utilize Hoechst 33258 fluorescent dye to the detection method of active o content in living cells |
| CN106905958A (en) * | 2017-03-21 | 2017-06-30 | 贵州大学 | A kind of fluorescence probe based on trans cucurbit(7)uril, preparation method and application |
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| Title |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113150770A (en) * | 2021-02-24 | 2021-07-23 | 武汉纺织大学 | Molecular assembly fluorescent probe and preparation method and application thereof |
| CN113150770B (en) * | 2021-02-24 | 2022-07-05 | 武汉纺织大学 | Molecular assembly fluorescent probe and preparation method and application thereof |
| CN113670867A (en) * | 2021-07-16 | 2021-11-19 | 贵州大学 | Supermolecule fluorescence sensing array and method for detecting multiple metal ions in aqueous solution |
| CN113670867B (en) * | 2021-07-16 | 2023-08-01 | 贵州大学 | Supermolecule fluorescent sensing array and method for detecting multiple metal ions in aqueous solution |
| CN113880851A (en) * | 2021-11-12 | 2022-01-04 | 广东科学技术职业学院 | Trifluorene bridged hexaimidazole macrocyclic compound and preparation method and application thereof |
| CN113880851B (en) * | 2021-11-12 | 2022-07-29 | 广东科学技术职业学院 | Trifluorene bridged hexaimidazole macrocyclic compound and preparation method and application thereof |
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